Additional file 3: Figure S3. of Epidermal growth factor receptor inhibitor with fluorouracil, leucovorin, and irinotecan as an alternative treatment for advanced upper tract urothelial carcinoma: a case report

Electropherogram of exon 2 of the KRAS gene. (TIF 3851 kb

Additional file 2: Figure S2. of Epidermal growth factor receptor inhibitor with fluorouracil, leucovorin, and irinotecan as an alternative treatment for advanced upper tract urothelial carcinoma: a case report

Electropherogram of the telomerase reverse transcriptase (TERT) promoter region. (TIF 3855 kb

Additional file 1: Figure S1. of Epidermal growth factor receptor inhibitor with fluorouracil, leucovorin, and irinotecan as an alternative treatment for advanced upper tract urothelial carcinoma: a case report

(A) Photomicrograph of the rectum showed the presence of neoplastic cells resembling adenocarcinoma (arrow). (B) Microscopic features of renal pelvis revealed infiltrating urothelial carcinoma, high grade (arrow). (C) Microscopic features of ureter revealed infiltrating urothelial carcinoma, high grade (arrow). (D) Microscopic features of bladder revealed infiltrating urothelial carcinoma, high grade (arrow). A–D show hematoxylin and eosin stain, magnification ×200. (TIF 2307 kb

Immunofluorescence staining results for detecting activated microglia expression in the spinal cord ventral horn.

<p>(A) Increased expression of activated microglia was observed in Group B compared with Group A. EPO administration resulted in lower microglia expression in Groups C and D compared with Group B (scale bars: 100 μm). (B) Quantitative analysis of activated microglia revealed a significant increase in the number of activated microglia in Group B compared with Group A. EPO treatment attenuated microglia activation. Values are expressed as a percentage of the mean ± SEM (n = 6). *: p < 0.05; **: p < 0.01.</p

Immunofluorescence staining images and quantitative analysis of COX-2 and iNOS in the spinal cord ventral horn.

<p>The immunofluorescence staining of COX-2 (A) and iNOS (B) 7 days after burn injury. The quantitative analysis of COX-2 (C) and iNOS (D) from spinal cord tissues in each experimental group has been assessed. COX-2 and iNOS expression decreased in the EPO-treated groups (Groups C and D) compared with the untreated group (Group B). *: p < 0.05; **: p < 0.01.</p

Grouping and flow chart of the animal study procedure.

<p>Sham burn or burn injury was induced at day 0. Following the procedure, wound care with 1% SSD was repeated twice a day. EPO was given for pharmacological investigation in Group C and D with different regimens. After completion of the protocol, all rats were sacrificed on week 1 (W1). Spinal cord ventral horn and gastrocnemius muscles were taken and processed by histopathological examination or Western blotting. Abbreviation: D, day; W, week.</p

Effect of EPO on hematological parameters.

<p>Effect of EPO on hematological parameters.</p

EPO alleviated burn-induced muscle cells apoptosis by Western blotting.

<p>Burn injury significantly increased the expression of cleaved caspase-3 in muscle cells vs Group A (p < 0.01), and Groups C and D exhibited decreased cleaved caspase-3 expression (p < 0.05) vs Group B.</p

EPO attenuates burn-induced motor neruon apoptosis.

<p>(A) Expression levels of BCL-2, BAX, and cleaved caspase-3 in the spinal cord ventral horn, measured through Western blotting. (B) Significant reduction in BCL-2/BAX expression was observed in Group B compared with Group A (100%). However, BCL-2/BAX expression was upregulated after EPO treatment. (C) Cleaved caspase-3 expression increased in the burn-induced group compared with the sham-control group. Similar to the notable reduction in cleaved caspase-3 expression following EPO treatment, a decrease in cleaved caspase-3 was observed. *: p < 0.05.</p

The Effects of EPO on autophagy markers in burn injury model by immunofluorescence analysis.

<p>(A, B) Double immunofluorescence staining and merged images using LC3B, ATG5, and NeuN in the spinal cord ventral horn were shown. The quantitative analysis of ATG5 (C) and LC3B activity (D) were measured. Burn injury significantly increased LC3B and ATG5 immunoreactivity in Group B versus Group A. After EPO treatment, LC3B and ATG5 immunoreactivity decreased markedly in Groups C and D vs Group B (**: p < 0.01; *: p < 0.05 versus Group B; Group A: 100%). Scale bars: 50 μm.</p